Search results for: resistant microbial strains
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2554

Search results for: resistant microbial strains

2104 Temperature Dependence and Seasonal Variation of Denitrifying Microbial Consortia from a Woodchip Bioreactor in Denmark

Authors: A. Jéglot, F. Plauborg, M. K. Schnorr, R. S. Sørensen, L. Elsgaard

Abstract:

Artificial wetlands such as woodchip bioreactors are efficient tools to remove nitrate from agricultural wastewater with a minimized environmental impact. However, the temperature dependence of the microbiological nitrate removal prevents the woodchip bioreactors from being an efficient system when the water temperature drops below 8℃. To quantify and describe the temperature effects on nitrate removal efficiency, we studied nitrate-reducing enrichments from a woodchip bioreactor in Denmark based on samples collected in Spring and Fall. Growth was quantified as optical density, and nitrate and nitrous oxide concentrations were measured in time-course experiments to compare the growth of the microbial population and the nitrate conversion efficiencies at different temperatures. Ammonia was measured to indicate the importance of dissimilatory nitrate reduction to ammonia (DNRA) in nitrate conversion for the given denitrifying community. The temperature responses observed followed the increasing trend proposed by the Arrhenius equation, indicating higher nitrate removal efficiencies at higher temperatures. However, the growth and the nitrous oxide production observed at low temperature provided evidence of the psychrotolerance of the microbial community under study. The assays conducted showed higher nitrate removal from the microbial community extracted from the woodchip bioreactor at the cold season compared to the ones extracted during the warmer season. This indicated the ability of the bacterial populations in the bioreactor to evolve and adapt to different seasonal temperatures.

Keywords: agricultural waste water treatment, artificial wetland, denitrification, psychrophilic conditions

Procedia PDF Downloads 102
2103 Damage Strain Analysis of Parallel Fiber Eutectic

Authors: Jian Zheng, Xinhua Ni, Xiequan Liu

Abstract:

According to isotropy of parallel fiber eutectic, the no- damage strain field in parallel fiber eutectic is obtained from the flexibility tensor of parallel fiber eutectic. Considering the damage behavior of parallel fiber eutectic, damage variables are introduced to determine the strain field of parallel fiber eutectic. The damage strains in the matrix, interphase, and fiber of parallel fiber eutectic are quantitatively analyzed. Results show that damage strains are not only associated with the fiber volume fraction of parallel fiber eutectic, but also with the damage degree.

Keywords: damage strain, initial strain, fiber volume fraction, parallel fiber eutectic

Procedia PDF Downloads 546
2102 Characterisation of Extracellular Polymeric Substances from Bacteria Isolated from Acid Mine Decant in Gauteng, South Africa

Authors: Nonhlanhla Nkosi, Kulsum Kondiah

Abstract:

The toxicological manifestation of heavy metals motivates interest towards the development of a reliable, eco-friendly biosorption process. With that being said, the aim of the current study was to characterise the EPS from heavy-metal resistant bacteria isolated from acid mine decant on the West Rand, Gauteng, South Africa. To achieve this, six exopolysaccharide (EPS) producing, metal resistant strains (Pb101, Pb102, Pb103, Pb204, Co101, and Ni101) were identified as Bacillus safensis strain NBRC 100820, Bacillus proteolyticus, Micrococcus luteus, Enterobacter sp. Pb204, Bacillus wiedmannii and Bacillus zhangzhouensis, respectively with 16S rRNA sequencing. Thereafter, EPS was extracted using chemical (formaldehyde/NaOH) and physical (ultrasonification) methods followed by physicochemical characterisation of carbohydrate, DNA, and protein contents using chemical assays and spectroscopy (FTIR- Fourier transformed infrared and 3DEEM- three-dimensional excitation-emission matrix fluorescence spectroscopy). EPS treated with formaldehyde/NaOH showed better recovery of macromolecules than ultrasonification. The results of the present study showed that carbohydrates were more abundant than proteins, with carbohydrate and protein concentrations of 8.00 mg/ml and 0.22 mg/ml using chemical method in contrast to 5.00 mg/ml and 0.77 mg/ml using physical method, respectively. The FTIR spectroscopy results revealed that the extracted EPS contained hydroxyl, amide, acyl, and carboxyl groups that corresponded to the aforementioned chemical analysis results, thus asserting the presence of carbohydrates, DNA, polysaccharides, and proteins in the EPS. These findings suggest that identified functional groups of EPS form surface charges, which serve as the binding sites for suspended particles, thus possibly mediating adsorption of divalent cations and heavy metals. Using the extracted EPS in the development of a cost-effective biosorption solution for industrial wastewater treatment is attainable.

Keywords: biosorbent, exopolysaccharides, heavy metals, wastewater treatment

Procedia PDF Downloads 134
2101 Antibacterial Activity of Bacillus thuringiensis Activated Delta-endotoxins

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

Abstract:

For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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2100 Effect of Fertilization and Combined Inoculation with Azospirillum brasilense and Pseudomonas fluorescens on Rhizosphere Microbial Communities of Avena sativa (Oats) and Secale Cereale (Rye) Grown as Cover Crops

Authors: Jhovana Silvia Escobar Ortega, Ines Eugenia Garcia De Salamone

Abstract:

Cover crops are an agri-technological alternative to improve all properties of soils. Cover crops such as oats and rye could be used to reduce erosion and favor system sustainability when they are grown in the same agricultural cycle of the soybean crop. This crop is very profitable but its low contribution of easily decomposable residues, due to its low C/N ratio, leaves the soil exposed to erosive action and raises the need to reduce its monoculture. Furthermore, inoculation with the plant growth promoting rhizobacteria contributes to the implementation, development and production of several cereal crops. However, there is little information on its effects on forage crops which are often used as cover crops to improve soil quality. In order to evaluate the effect of combined inoculation with Azospirillum brasilense and Pseudomonas fluorescens on rhizosphere microbial communities, field experiments were conducted in the west of Buenos Aires province, Argentina, with a split-split plot randomized complete block factorial design with three replicates. The factors were: type of cover crop, inoculation and fertilization. In the main plot two levels of fertilization 0 and 7 40-0-5 (NPKS) were established at sowing. Rye (Secale cereale cultivar Quehué) and oats (Avena sativa var Aurora.) were sown in the subplots. In the sub-subplots two inoculation treatments are applied without and with application of a combined inoculant with A. brasilense and P. fluorescens. Due to the growth of cover crops has to be stopped usually with the herbicide glyphosate, rhizosphere soil of 0-20 and 20-40 cm layers was sampled at three sampling times which were: before glyphosate application (BG), a month after glyphosate application (AG) and at soybean harvest (SH). Community level of physiological profiles (CLPP) and Shannon index of microbial diversity (H) were obtained by multivariate analysis of Principal Components. Also, the most probable number (MPN) of nitrifiers and cellulolytics were determined using selective liquid media for each functional group. The CLPP of rhizosphere microbial communities showed significant differences between sampling times. There was not interaction between sampling times and both, types of cover crops and inoculation. Rhizosphere microbial communities of samples obtained BG had different CLPP with respect to the samples obtained in the sampling times AG and SH. Fertilizer and depth of sampling also caused changes in the CLPP. The H diversity index of rhizosphere microbial communities of rye in the sampling time BG were higher than those associated with oats. The MPN of both microbial functional types was lower in the deeper layer since these microorganisms are mostly aerobic. The MPN of nitrifiers decreased in rhizosphere of both cover crops only AG. At the sampling time BG, the NMP of both microbial types were larger than those obtained for AG and SH. This may mean that the glyphosate application could cause fairly permanent changes in these microbial communities which can be considered bio-indicators of soil quality. Inoculation and fertilizer inputs could be included to improve management of these cover crops because they can have a significant positive effect on the sustainability of the agro-ecosystem.

Keywords: community level of physiological profiles, microbial diversity, plant growth promoting rhizobacteria, rhizosphere microbial communities, soil quality, system sustainability

Procedia PDF Downloads 378
2099 Post-Harvest Biopreservation of Fruit and Vegetables with Application of Lactobacillus Strains

Authors: Judit Perjessy, Zsolt Zalan, Ferenc Hegyi, Eniko Horvath-Szanics, Krisztina Takacs, Andras Nagy, Adel Klupacs, Erika Koppany-Szabo, Zhirong Wang, Kaituo Wang, Muying Du, Jianquan Kan

Abstract:

The post-harvest diseases cause great economic losses in the fruit and vegetables; the prevention of these deterioration has great importance. Against the fungi, which cause most of the diseases, are extensively used the fungicides. However, there are increasing consumer concerns over the presence of pesticide residues in food. An alternative and in recent years, increasingly studied method for the prevention of the diseases is biocontrol, where antagonistic microorganisms are used for the control of fungi. The genera of Lactobacillus is well known and extensively studied, but its applicability as biocontrol agents in post-harvest preservation of fruit and vegetables is poorly investigated. However these bacteria can be found on the surface of the plants and have great antimicrobial activity. In our study we have investigated the chitinase activity, the antifungal effect and the applicability of several Lactobacillus strains to select potential biocontrol agents. We investigated the determination of the environmental parameters of a gene (encoding chitinase) expression and we also investigated the relationship between actual antifungal activity and potential chitinase activity. Mixed cultures were also developed to enhance the antifungal activity and determined the optimal mold spore and bacteria concentration ratio for the appropriate efficacy. Five Lactobacillus strains (L. acidophilus N2, L. delbrueckii subsp. bulgaricus B397, L. sp. 2231, L. sake subsp. sake 2471, L. buchneri 1145) possess chitinase-coding gene from the 43 investigated Lactobacillus strains. Proteins with similar molecular weight and separation properties like bacterial chitinases were detected from these strains, which also possess chitin-binding property. Nevertheless, they were inactive, lacks the chitinolytic activity. In point of the cumulative activity of inhibition, our results showed that certain strains were statistically significant in a positive direction compared to other strains, e.g., L. rhamnosus VT1 and L. Casey 154 have shown great general antifungal effect against 11 molds from the genera Penicillium and Botrytis and isolated from spoiled fruit and vegetables. Also, some mixed cultures (L. rhamnosus VT1 - L. Plantarum 299v) showed significant antifungal effects against the indigenous molds on the surface of apple fruit during the industrial storage experiment. Thus, they could be promising for post-harvest biopreservation.

Keywords: biocontrol, chitinase, Lactobacillus, post-harvest

Procedia PDF Downloads 137
2098 Environmental Health Risk Assessment of Hospital Wastewater in Enugu Urban, Nigeria

Authors: C. T. Eze, I. N. E. Onwurah

Abstract:

An important hydrogeologic problem in areas of high faults formations is high environmental health hazard occasioned by microbial and heavy metals contamination of ground waters. Consequently, we examined the microbial load and heavy metals concentration of hospital wastewater discharged into the environment at Park Lane General Hospital Enugu Urban, Nigeria. The microbial counts, characteristics and frequency of occurrences of the isolated microorganisms were determined by cultural, morphological and biochemical characteristics using established procedure while the varying concentrations of the identified heavy metals were determined using the spectrophotometric method. The microbiological analyses showed a mean total aerobic bacteria counts from 13.7 ± 0.65 × 107 to 22.8 ± 1.14 ×1010 CFU/ml, mean total anaerobic bacteria counts from 6.0 ± 1.6 × 103 to 1.7 ± 0.41 ×104 CFU/ml and mean total fungal counts from 0 ± 0 to 2.3 ± 0.16 × 105 CFU/ml. The isolated micro-organisms which included both pathogenic and non-pathogenic organisms were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Bacillus subtilis, Proteus vulgaris, Klesbsiella pneumonia and bacteriodes sp. The only fungal isolate was Candida albican. The heavy metals identified in the leachate were Arsenic, Cadmium, Lead, Mercury and Chromium and their concentrations ranged from 0.003 ± 0.00082 to 0.14 ± 0.0082 mg/l. These values were above WHO permissible limits while others fall within the limits. Therefore, hospital waste water can pose the environmental health risk when not properly treated before discharge, especially in geologic formations with high fault formations.

Keywords: bacterial isolates, fungal isolates, heavy metals, hospital wastewater, microbial counts

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2097 Survival of Four Probiotic Strains in Acid, Bile Salt and After Spray Drying

Authors: Rawichar Chaipojjana, Suttipong Phosuksirikul, Arunsri Leejeerajumnean

Abstract:

The objective of the study was to select the survival of probiotic strains when exposed to acidic and bile salts condition. Four probiotic strains (Lactobacillus casei subsp. rhamnosus TISTR 047, Lactobacillus casei TISTR 1500, Lactobacillus acidophilus TISTR 1338 and Lactobacillus plantarum TISTR 1465) were cultured in MRS broth and incubated at 35ºC for 15 hours before being inoculated into acidic condition (5 M HCl, pH 2) for 2 hours and bile salt (0.3%, pH 5.8) for 8 hour. The survived probiotics were counted in MRS agar. Among four stains, Lactobacillus casei subsp. rhamnosus TISTR 047 was the highest tolerance specie. Lactobacillus casei subsp. rhamnosus TISTR 047 reduced 6.74±0.07 log CFU/ml after growing in acid and 5.52±0.05 log CFU/ml after growing in bile salt. Then, double emulsion of microorganisms was chosen to encapsulate before spray drying. Spray drying was done with the inlet temperature 170ºC and outlet temperature 80ºC. The results showed that the survival of encapsulated Lactobacillus casei subsp. rhamnosus TISTR 047 after spray drying decreased from 9.63 ± 0.32 to 8.31 ± 0.11 log CFU/ml comparing with non-encapsulated, 9.63 ± 0.32 to 4.06 ± 0.08 log CFU/ml. Therefore, Lactobacillus casei subsp. rhamnosus TISTR 047 would be able to survive in gastrointestinal and spray drying condition.

Keywords: probiotic, acid, bile salt, spray drying

Procedia PDF Downloads 338
2096 Antibacterial Activity of Bacillus thuringiensis Cristalline Parasporal Proteins

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

Abstract:

For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

Procedia PDF Downloads 416
2095 Development of an in vitro Fermentation Chicken Ileum Microbiota Model

Authors: Bello Gonzalez, Setten Van M., Brouwer M.

Abstract:

The chicken small intestine represents a dynamic and complex organ in which the enzymatic digestion and absorption of nutrients take place. The development of an in vitro fermentation chicken small intestinal model could be used as an alternative to explore the interaction between the microbiota and nutrient metabolism and to enhance the efficacy of targeting interventions to improve animal health. In the present study we have developed an in vitro fermentation chicken ileum microbiota model for unrevealing the complex interaction of ileum microbial community under physiological conditions. A two-vessel continuous fermentation process simulating in real-time the physiological conditions of the ileum content (pH, temperature, microaerophilic/anoxic conditions, and peristaltic movements) has been standardized as a proof of concept. As inoculum, we use a pool of ileum microbial community obtained from chicken broilers at the age of day 14. The development and validation of the model provide insight into the initial characterization of the ileum microbial community and its dynamics over time-related to nutrient assimilation and fermentation. Samples can be collected at different time points and can be used to determine the microbial compositional structure, dynamics, and diversity over time. The results of studies using this in vitro model will serve as the foundation for the development of a whole small intestine in vitro fermentation chicken gastrointestinal model to complement our already established in vitro fermentation chicken caeca model. The insight gained from this model could provide us with some information about the nutritional strategies to restore and maintain chicken gut homeostasis. Moreover, the in vitro fermentation model will also allow us to study relationships between gut microbiota composition and its dynamics over time associated with nutrients, antimicrobial compounds, and disease modelling.

Keywords: broilers, in vitro model, ileum microbiota, fermentation

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2094 Ficus Microcarpa Fruit Derived Iron Oxide Nanomaterials and Its Anti-bacterial, Antioxidant and Anticancer Efficacy

Authors: Fuad Abdullah Alatawi

Abstract:

Microbial infections-based diseases are a significant public health issue around the world, mainly when antibiotic-resistant bacterium types evolve. In this research, we explored the anti-bacterial and anti-cancer potency of iron-oxide (Fe₂O₃) nanoparticles prepared from F. macrocarpa fruit extract. The chemical composition of F. macrocarpa fruit extract was used as a reducing and capping agent for nanoparticles’ synthesis was examined by GC-MS/MS analysis. Then, the prepared nanoparticles were confirmed by various biophysical techniques, including X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), UV-Vis Spectroscopy, and Transmission Electron Microscopy (TEM) and Energy Dispersive Spectroscopy (EDAX), and Dynamic Light Scattering (DLS). Also, the antioxidant capacity of fruit extract was determined through 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), Fluorescence Recovery After Photobleaching (FRAP), Superoxide Dismutase (SOD) assays. Furthermore, the cytotoxicity activities of Fe₂O₃ NPs were determined using the (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (MTT) test on MCF-7 cells. In the antibacterial assay, lethal doses of the Fe₂O₃NPs effectively inhibited the growth of gram-negative and gram-positive bacteria. The surface damage, ROS production, and protein leakage are the antibacterial mechanisms of Fe₂O₃NPs. Concerning antioxidant activity, the fruit extracts of F. macrocarpa had strong antioxidant properties, which were confirmed by DPPH, ABTS, FRAP, and SOD assays. In addition, the F. microcarpa-derived iron oxide nanomaterials greatly reduced the cell viability of (MCF-7). The GC-MS/MS analysis revealed the presence of 25 main bioactive compounds in the F. microcarpa extract. Overall, the finding of this research revealed that F. microcarpa-derived Fe₂O₃ nanoparticles could be employed as an alternative therapeutic agent to cure microbial infection and breast cancer in humans.

Keywords: ficus microcarpa, iron oxide, antibacterial activity, cytotoxicity

Procedia PDF Downloads 97
2093 Toxicological Standardization of Heavy Metals and Microbial Contamination Haematinic Herbal Formulations Marketed in India

Authors: A. V. Chandewar, Sanjay Bais

Abstract:

Backgound: In India, drugs of herbal origin have been used in traditional systems of medicines such as Unani and Ayurveda since ancient times. WHO limit for Escherichia coli is 101/gm cfu, for Staphylococus aureus 105/gm cfu, and for Pseudomonas aeruginosa 103/gm cfu and for Salmonella species nil cfu. WHO mentions maximum permissible limits in raw materials only for arsenic, cadmium, and lead, which amount to 1.0, 0.3, and 10 ppm, respectively. Aim: The main purpose of the investigation was to document evidence for the users, and practitioners of marketed haematinic herbal formulations. In the present study haematinic herbal formulations marketed in Yavatmal India were determined for the presence of microbial and heavy metal content. Method: The investigations were performed by using specific medias and atomic absorption spectrometry. Result: The present work indicates the presence of heavy metal contents in herbal formulations selected for study. It was found that arsenic content in formulations was below the permissible limit in all formulations. The cadmium and lead content in six formulations were above the permissible limits. Such formulations are injurious to health of patient if consumed regularly. The specific medias were used to determining the presence of Escherichia coli 4 samples, Staphylococcus aureus 3 samples, and P. aeruginosa 4 samples. The data indicated suggest that there is requirement of in process improvement to provide better quality for consumer health in order to be competitive in international markets. Summary/Conclusion: The presence of microbial and heavy metal content above WHO limits indicates that the GMP was not followed during manufacturing of herbal formulations marketed in India.

Keywords: toxicological standardization, heavy metals, microbial contamination, haematinic herbal formulations

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2092 Antimicrobial Activity of Different Essential Oils in Synergy with Amoxicillin against Clinical Isolates of Methicillin-Resistant Staphylococcus aureus

Authors: Naheed Niaz, Nimra Naeem, Bushra Uzair, Riffat Tahira

Abstract:

Antibacterial activity of different traditional plants essential oils against clinical isolates of Methicillin-resistant Staphylococcus aureus (MRSA) through disk diffusion method was evaluated. All the tested essential oils, in different concentrations, inhibited growth of S. aureus to varying degrees. Cinnamon and Thyme essential oils were observed to be the “best” against test pathogen. Even at lowest concentration of these essential oils i.e. 25 µl/ml, clear zone of inhibition was recorded 9+0.085mm and 8+0.051mm respectively, and at higher concentrations there was a total reduction in growth of MRSA. The study also focused on analyzing the synergistic effects of essential oils in combination with amoxicillin. Results showed that oregano and pennyroyal mint essential oils which were not very effective alone turned out to be strong synergistic enhancers. The activity increased with increase in concentration of the essential oils. It may be concluded from present results that cinnamon and thyme essential oils could be used as potential antimicrobial source for the treatment of infections caused by Methicillin-resistant Staphylococcus aureus (MRSA).

Keywords: Staphylococcus aureus, essential oils, antibiotics, combination therapy, minimum inhibitory concentration

Procedia PDF Downloads 426
2091 Isolation and Selection of Strains Perspective for Sewage Sludge Processing

Authors: A. Zh. Aupova, A. Ulankyzy, A. Sarsenova, A. Kussayin, Sh. Turarbek, N. Moldagulova, A. Kurmanbayev

Abstract:

One of the methods of organic waste bioconversion into environmentally-friendly fertilizer is composting. Microorganisms that produce hydrolytic enzymes play a significant role in accelerating the process of organic waste composting. We studied the enzymatic potential (amylase, protease, cellulase, lipase, urease activity) of bacteria isolated from the sewage sludge of Nur-Sultan, Rudny, and Fort-Shevchenko cities, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha for processing organic waste and identifying active strains. Microorganism isolation was carried out by the cultures enrichment method on liquid nutrient media, followed by inoculating on different solid media to isolate individual colonies. As a result, sixty-one microorganisms were isolated, three of which were thermophiles (DS1, DS2, and DS3). The highest number of isolates, twenty-one and eighteen, were isolated from sewage sludge of Nur-Sultan and Rudny cities, respectively. Ten isolates were isolated from the wastewater of the sewage treatment plant in Fort-Shevchenko. From the dacha soil of Nur-Sultan city and freshly cut grass - 9 and 5 isolates were revealed, respectively. The lipolytic, proteolytic, amylolytic, cellulolytic, ureolytic, and oil-oxidizing activities of isolates were studied. According to the results of experiments, starch hydrolysis (amylolytic activity) was found in 2 isolates - CB2/2, and CB2/1. Three isolates - CB2, CB2/1, and CB1/1 were selected for the highest ability to break down casein. Among isolated 61 bacterial cultures, three isolates could break down fats - CB3, CBG1/1, and IL3. Seven strains had cellulolytic activity - DS1, DS2, IL3, IL5, P2, P5, and P3. Six isolates rapidly decomposed urea. Isolate P1 could break down casein and cellulose. Isolate DS3 was a thermophile and had cellulolytic activity. Thus, based on the conducted studies, 15 isolates were selected as a potential for sewage sludge composting - CB2, CB3, CB1/1, CB2/2, CBG1/1, CB2/1, DS1, DS2, DS3, IL3, IL5, P1, P2, P5, P3. Selected strains were identified on a mass spectrometer (Maldi-TOF). The isolate - CB 3 was referred to the genus Rhodococcus rhodochrous; two isolates CB2 and CB1 / 1 - to Bacillus cereus, CB 2/2 - to Cryseobacterium arachidis, CBG 1/1 - to Pseudoxanthomonas sp., CB2/1 - to Bacillus megaterium, DS1 - to Pediococcus acidilactici, DS2 - to Paenibacillus residui, DS3 - to Brevibacillus invocatus, three strains IL3, P5, P3 - to Enterobacter cloacae, two strains IL5, P2 - to Ochrobactrum intermedium, and P1 - Bacillus lichenoformis. Hence, 60 isolates were isolated from the wastewater of the cities of Nur-Sultan, Rudny, Fort-Shevchenko, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha. Based on the highest enzymatic activity, 15 active isolates were selected and identified. These strains may become the candidates for bio preparation for sewage sludge processing.

Keywords: sewage sludge, composting, bacteria, enzymatic activity

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2090 Development of Low-Cost Vibro-Acoustic, and Fire-Resistant, Insulation Material from Natural and Sustainable Sources

Authors: K. Nasir, S. Ahmad, A. Khan, H. Benkreira

Abstract:

The topic of the research is to develop sustainable fire-resistant materials for vibration and acoustic damping of structure and airborne noises from sustainable recycled materials and biodegradable binders. The paper reports, methods and techniques of enhancing fire resistive, vibration and acoustic properties of building insulation materials made from natural resources like wood and recycled materials like rubber and textile waste. The structures are designed to optimize the number, size and stratification of closed (heat insulating) and open (noise insulating) pores. The samples produced are tested for their heat and noise insulating properties, including vibration damping and their structural properties (airflow resistivity, porosity, tortuosity and elastic modulus). The structural properties are then used in theoretical models to check the acoustic insulation measurements. Initial data indicate that one layer of such material can yield as much as 18 times more damping, increasing the loss factor by 18%.

Keywords: fire resistant, vibration damping, acoustic material, vibro-acoustic, thermal insulation, sustainable material, low cost materials, recycled materials, construction material

Procedia PDF Downloads 113
2089 The Use of Ketamine in Conjunction with Antidepressants for Treatment Resistant Depression

Authors: Zumra Mehmedovic, Susan Luhrmann

Abstract:

Treatment-resistant depression (TRD) is a debilitating mental health disorder for which there are very few available treatment options. Current research suggests that ketamine may be a safe and effective option for the treatment of TRD. Research utilizing a review of the literature was conducted to determine if ketamine in conjunction with antidepressants is more effective than antidepressants alone in the treatment of TRD. The literature consists of ten journal articles which include quantitative studies based on primary research. A critique of the literature was done to determine whether the findings are reliable, critiquing elements influencing the believability and robustness of the research. The research was based on the neuroplasticity theory of depression, hypothesizing that ketamine, in conjunction with antidepressants, will be more effective than antidepressants alone as they have different mechanisms of action. All the studies except one found ketamine in conjunction with antidepressants to be a more effective treatment than antidepressants alone in the treatment of TRD. Results of the studies indicate that ketamine is effective in treating TRD at various doses, settings, and routes of administration. Further research is necessary, though, to further explore and confirm the findings. Several gaps in literature were identified, including the optimal dose of ketamine, its long-term efficacy and safety, and effects of ketamine in repeated doses. The research topic is highly significant to advanced practice nursing, as based on the findings, ketamine can be utilized as a safe and effective treatment for TRD.

Keywords: ketamine, major depressive disorder, treatment-resistant depression, treatment

Procedia PDF Downloads 113
2088 Settings of Conditions Leading to Reproducible and Robust Biofilm Formation in vitro in Evaluation of Drug Activity against Staphylococcal Biofilms

Authors: Adela Diepoltova, Klara Konecna, Ondrej Jandourek, Petr Nachtigal

Abstract:

A loss of control over antibiotic-resistant pathogens has become a global issue due to severe and often untreatable infections. This state is reflected in complicated treatment, health costs, and higher mortality. All these factors emphasize the urgent need for the discovery and development of new anti-infectives. One of the most common pathogens mentioned in the phenomenon of antibiotic resistance are bacteria of the genus Staphylococcus. These bacterial agents have developed several mechanisms against the effect of antibiotics. One of them is biofilm formation. In staphylococci, biofilms are associated with infections such as endocarditis, osteomyelitis, catheter-related bloodstream infections, etc. To author's best knowledge, no validated and standardized methodology evaluating candidate compound activity against staphylococcal biofilms exists. However, a variety of protocols for in vitro drug activity testing has been suggested, yet there are often fundamental differences. Based on our experience, a key methodological step that leads to credible results is to form a robust biofilm with appropriate attributes such as firm adherence to the substrate, a complex arrangement in layers, and the presence of extracellular polysaccharide matrix. At first, for the purpose of drug antibiofilm activity evaluation, the focus was put on various conditions (supplementation of cultivation media by human plasma/fetal bovine serum, shaking mode, the density of initial inoculum) that should lead to reproducible and robust in vitro staphylococcal biofilm formation in microtiter plate model. Three model staphylococcal reference strains were included in the study: Staphylococcus aureus (ATCC 29213), methicillin-resistant Staphylococcus aureus (ATCC 43300), and Staphylococcus epidermidis (ATCC 35983). The total biofilm biomass was quantified using the Christensen method with crystal violet, and results obtained from at least three independent experiments were statistically processed. Attention was also paid to the viability of the biofilm-forming staphylococcal cells and the presence of extracellular polysaccharide matrix. The conditions that led to robust biofilm biomass formation with attributes for biofilms mentioned above were then applied by introducing an alternative method analogous to the commercially available test system, the Calgary Biofilm Device. In this test system, biofilms are formed on pegs that are incorporated into the lid of the microtiter plate. This system provides several advantages (in situ detection and quantification of biofilm microbial cells that have retained their viability after drug exposure). Based on our preliminary studies, it was found that the attention to the peg surface and substrate on which the bacterial biofilms are formed should also be paid to. Therefore, further steps leading to the optimization were introduced. The surface of pegs was coated by human plasma, fetal bovine serum, and L-polylysine. Subsequently, the willingness of bacteria to adhere and form biofilm was monitored. In conclusion, suitable conditions were revealed, leading to the formation of reproducible, robust staphylococcal biofilms in vitro for the microtiter model and the system analogous to the Calgary biofilm device, as well. The robustness and typical slime texture could be detected visually. Likewise, an analysis by confocal laser scanning microscopy revealed a complex three-dimensional arrangement of biofilm forming organisms surrounded by an extracellular polysaccharide matrix.

Keywords: anti-biofilm drug activity screening, in vitro biofilm formation, microtiter plate model, the Calgary biofilm device, staphylococcal infections, substrate modification, surface coating

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2087 Effect of Ethanol and Betadine on the Preformed Biofilm of Staphylococcus Aureus Isolated from Urinary Catheter

Authors: Kara Terki Ibtissem, Hassaine Hafida, Bellifa Samia

Abstract:

Introduction: Staphylococcus aureus is one of the species that are most frequently isolated from urinary catheters. The ability to produce a biofilm is an important step in the pathogenesis of these staphylococci; biofilm formation is strongly dependent on the environmental conditions it also depends on the different parameters these biofilms are subjected to. Antiseptics, including ethanol and betadine, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that disinfectants may enhance biofilm production in Staphylococci. Methods: In this study, 48 staphylococcus aureus isolated from urinary catheters at the University Hospital Center of Sidi Bel Abbes (in Northwestern Algeria) were analyzed to detect the formation of biofilm by culture on Red Congo Agar (RCA), the Tube Method (TM) and tissue Culture Plate (TCP) techniques, this last was also used to investigate the effect of ethanol and Betadine on the preformed biofilm In a second time to know which environment is most favorable to the formation of the biofilm we perform a statistical test based on the student test by the software R. Results: It has been found that 23 strains produced a bacterial slime on the Congo red medium, 5 strains produced a biofilm by the tube method, 2 of which are highly productive. In addition, 7 strains produced a biofilm on polystyrene micro-plates; this number was higher in the presence of ethanol 70% and ethanol 90% with 19 and 11 biofilm-producing strains, respectively. On the other hand, no biofilm was formed in the presence of Betadine. Conclusion: It is important to examine the response of biofilms following an imposed external constraint, such as disinfectants, in order to develop new strategies to combat bacterial biofilms but also to better control their formation.

Keywords: staphylococcus aureus, biofilm, urinary catheter, ethanol

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2086 Absence of Vancomycin-Resistant Enterococci Amongst Urban and Rural Hooded Crows in Hungary

Authors: Isma Benmazouz, Bálint Joszef Nagy, Bence Bálacs, Gábor Kardos, László Kővér

Abstract:

Vancomycin-resistant enterococci (VRE) are among the major nosocomial threats, which have a potential for zoonotic transmission due to the ubiquity of enterococci in the environment and in animal microbiota, e.g., wild birds. . In order to assess the prevalence in an urbanized bird species, 221 fecal samples were collected from Hooded crows (Corvus cornix) in 2020. Fecal samples were screened using VRE agar plates. None of the samples yielded VRE. The absence of VRE isolates in sampled urban hooded crows indicates that crows residing in the city do not necessarily constitute a reservoir of VREs.

Keywords: resistance, crows, Enterococci, wild birds

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2085 Expression Regulation of Membrane Protein by Codon Variation of Amino Acid at N-Terminal Region

Authors: Ahreum Choi, Otgontuya Tsogbadrakh, Kwang-Hwan Jung

Abstract:

Microbial rhodopsins are well-known seven-transmembrane proteins that have been extensively studied. These retinal-binding proteins have divided into two types. The type I is microbial rhodopsin, and type II (visual pigment) is expressed mostly in mammalian eyes. For type I rhodopsin, there are two main functions that are ion pumping activity and sensory transduction. Anabaena sensory rhodopsin (ASR) is one of the microbial rhodopsin with main function as photo-sensory transduction. Although ASR is expressed fairly well in Escherichia coli, the expression level is relatively less compare to Proteorhodopsin. In this study, full length of ASR was used to test for the expression influence by codon usage in E. coli. Eight amino acids of codon at N-terminal part of ASR were changed randomly with designed primers, which allow 8,192 nucleotide different cases. The codon changes were screened for the preferable codons of each residue, which have given higher expression yield. Among those 57 selected mutations, there are 24 color-enhanced E. coli colonies that contain ASR proteins, and it showed better expression level than the wild type ASR codon usage. This strongly suggests that high codon usage of only partial N-terminal of protein can increase the expression level of whole protein.

Keywords: 7-transmembrane, all-trans retinal, rhodopsin, codon-usage, protein expression

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2084 Applying Computer Simulation Methods to a Molecular Understanding of Flaviviruses Proteins towards Differential Serological Diagnostics and Therapeutic Intervention

Authors: Sergio Alejandro Cuevas, Catherine Etchebest, Fernando Luis Barroso Da Silva

Abstract:

The flavivirus genus has several organisms responsible for generating various diseases in humans. Special in Brazil, Zika (ZIKV), Dengue (DENV) and Yellow Fever (YFV) viruses have raised great health concerns due to the high number of cases affecting the area during the last years. Diagnostic is still a difficult issue since the clinical symptoms are highly similar. The understanding of their common structural/dynamical and biomolecular interactions features and differences might suggest alternative strategies towards differential serological diagnostics and therapeutic intervention. Due to their immunogenicity, the primary focus of this study was on the ZIKV, DENV and YFV non-structural proteins 1 (NS1) protein. By means of computational studies, we calculated the main physical chemical properties of this protein from different strains that are directly responsible for the biomolecular interactions and, therefore, can be related to the differential infectivity of the strains. We also mapped the electrostatic differences at both the sequence and structural levels for the strains from Uganda to Brazil that could suggest possible molecular mechanisms for the increase of the virulence of ZIKV. It is interesting to note that despite the small changes in the protein sequence due to the high sequence identity among the studied strains, the electrostatic properties are strongly impacted by the pH which also impact on their biomolecular interactions with partners and, consequently, the molecular viral biology. African and Asian strains are distinguishable. Exploring the interfaces used by NS1 to self-associate in different oligomeric states, and to interact with membranes and the antibody, we could map the strategy used by the ZIKV during its evolutionary process. This indicates possible molecular mechanisms that can explain the different immunological response. By the comparison with the known antibody structure available for the West Nile virus, we demonstrated that the antibody would have difficulties to neutralize the NS1 from the Brazilian strain. The present study also opens up perspectives to computationally design high specificity antibodies.

Keywords: zika, biomolecular interactions, electrostatic interactions, molecular mechanisms

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2083 Use of Soil Microorganisms for the Production of Electricity through Microbial Fuel Cells

Authors: Abhipsa Mohanty, Harit Jha

Abstract:

The world's energy demands are continuing to rise, resulting in a worldwide energy crisis and environmental pollution. Because of finite, declining supply and environmental damage, reliance on fossil fuels is unsustainable. As a result, experts are concentrating on alternative, renewable, and carbon-free energy sources. Energy sources that are both environmentally and economically sustainable are required. Microbial fuel cells (MFCs) have recently received a lot of attention due to their low operating temperatures and ability to use a variety of biodegradable substrates as fuel. There are single-chamber MFCs as well as traditional MFCs with anode and cathode compartments. Bioelectricity is produced when microorganisms actively catabolize substrate. MFCs can be used as a power source in small devices like biosensors. Understanding of its components, microbiological processes, limiting variables, and construction designs in MFC systems must be simplified, and large-scale systems must be developed for them to be cost-effective as well as increase electricity production. The purpose of this research was to review current microbiology knowledge in the field of electricity. The manufacturing process, the materials, and procedures utilized to construct the technology, as well as the applications of MFC technology, are all covered.

Keywords: bio-electricity, exoelectrogenic bacteria, microbial fuel cells, soil microorganisms

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2082 Antibacterial Activity of Trans-Cinnamaldehyde and Geraniol and Their Potential as Ingredients of Biocidal Polymers

Authors: Daria Olkiewicz, Maciej Walczak

Abstract:

In this paper, the biocidal effects of trans-cinnamaldehyde (a main component of cinnamon oil) and geraniol (a constituent of Pelargonium graveolens essential oil) are presented. The activity of the combination of trans-cinnamaldehyde and geraniol was tested against 3 bacterial strains: Staphylococcus aureus ATCC6538 (Gramm+), Escherichia coli ATCC8739 (Gramm-, Lac+) and Pseudomonas aeruginosa KKP 991(Gramm-, Lac-). The biocidal activity of trans-cinnamaldehyde-geraniol mixture against bacteria mentioned above was evaluated by disk-diffusion method. The model strains were exposed on 1, 2.5, 5 and 10 mg of trans-cinnamaldehyde-geraniol mixture per disk, and all strains were susceptible to this combination of plant compounds. For all microorganisms, also Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were estimated. For Staphylococcus aureus MIC was 0.0625 mg/ml of the trans-cinnamaldehyde and geraniol mixture, and MBC was 1.25 mg/ml; For Escherichia coli MIC=0.5 mg/ml, MBC=1 mg/ml, and finally Pseudomonas aeruginosa was inhibited in 0.5 mg/ml, and minimal biocidal concentration of tested mixture for it was 1.25 mg/ml. There are also reports about the synergistic working of trans-cinnamaldehyde and geraniol against microorganisms and the antimicrobial activity of polymers enriched with trans-cinnamaldehyde or geraniol, therefore the successful development and introduction to the today life of biocidal polymer enriched with trans-cinnamaldehyde and geraniol are possible.

Keywords: antibacterial activity, biocidal polymers, geraniol, trans-cinnamaldehyde

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2081 Effect of Sodium Alginate-based Edible Coating with Natural Essential Oils and Modified Atmosphere Packaging on Quality of Fresh-cut Pineapple

Authors: Muhammad Rafi Ullah Khan, Yaodong Guo, Vanee Chonhenchob, Jinjin Pei, Chongxing Huang

Abstract:

The effect of sodium alginate (1%) based edible coating incorporated natural essential oils; thymol, carvone and carvacrol as antimicrobial agents at different concentrations (0.1, 0.5 and 1.0 %) on the quality changes of fresh-cut pineapple were investigated. Pineapple dipped in distilled water was served as control. After coating, fruit were sealed in a modified atmosphere package (MAP) using high permeable film; and stored at 5 °C. Gas composition in package headspace, color values (L*, a*, b*, C*), TSS, pH, ethanol, browning, and microbial decay were monitored during storage. Oxygen concentration continuously decreased while carbon dioxide concentration inside all packages continuously increased over time. Color parameters (L*, b*, c*) decreased and a* values increased during storage. All essential oils significantly (p ≤ 0.05) prevented microbial growth than control. A significantly higher (p ≤ 0.05) ethanol content was found in the control than in all other treatments. Visible microbial growth, high ethanol, and low color values limited the shelf life to 6 days in control as compared to 9 days in all other treatments. Among all essential oils, thymol at all concentrations maintained the overall quality of the pineapple and could potentially be used commercially in fresh fruit industries for longer storage.

Keywords: essential oils, antibrowning agents, antimicrobial agents, modified atmosphere packaging, microbial decay, pineapple

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2080 Preparation and Characterization of Nanostructured FeN Electrocatalyst for Air Cathode Microbial Fuel Cell (MFC)

Authors: Md. Maksudur Rahman Khan, Chee Wai Woon, Huei Ruey Ong, Vignes Rasiah, Chin Kui Cheng, Kar Min Chan, E. Baranitharan

Abstract:

The present work represents a preparation of non-precious iron-based electrocatalyst (FeN) for ORR in air-cathode microbial fuel cell by pyrolysis treatment. Iron oxalate which recovered from the industrial wastewater and Phenanthroline (Phen) were used as the iron and nitrogen precursors, respectively in preparing FeN catalyst. The performance of as prepared catalyst (FeN) was investigated in a single chambered air cathode MFC in which anaerobic sludge was used as inoculum and palm oil mill effluent as substrate. The maximum open circuit potential (OCV) and the highest power density recorded were 0.543 V and 4.9 mW/m2, respectively. Physical characterization of FeN was elucidated by using Brunauner Emmett Teller (BET), X-Ray Diffraction (XRD) analysis and Field Emission Scanning Electron Microscopy (FESEM) while the electrochemical properties were characterized by cyclic voltammetry (CV) analysis. The presence of biofilm on anode surface was examined using FESEM and confirmed using Infrared Spectroscopy and Thermogravimetric Analysis. The findings of this study demonstrated that FeN is electrochemically active and further modification is needed to increase the ORR catalytic activity.

Keywords: iron based catalyst, microbial fuel cells, oxygen reduction reaction, palm oil mill effluent

Procedia PDF Downloads 306
2079 Some Extreme Halophilic Microorganisms Produce Extracellular Proteases with Long Lasting Tolerance to Ethanol Exposition

Authors: Cynthia G. Esquerre, Amparo Iris Zavaleta

Abstract:

Extremophiles constitute a potentially valuable source of proteases for the development of biotechnological processes; however, the number of available studies in the literature is limited compared to mesophilic counterparts. Therefore, in this study, Peruvian halophilic microorganisms were characterized to select suitable proteolytic strains that produce active proteases under exigent conditions. Proteolysis was screened using the streak plate method with gelatin or skim milk as substrates. After that, proteolytic microorganisms were selected for phenotypic characterization and screened by a semi-quantitative proteolytic test using a modified method of diffusion agar. Finally, proteolysis was evaluated using partially purified extracts by ice-cold ethanol precipitation and dialysis. All analyses were carried out over a wide range of NaCl concentrations, pH, temperature and substrates. Of a total of 60 strains, 21 proteolytic strains were selected, of these 19 were extreme halophiles and 2 were moderates. Most proteolytic strains demonstrated differences in their biochemical patterns, particularly in sugar fermentation. A total of 14 microorganisms produced extracellular proteases, 13 were neutral, and one was alkaline showing activity up to pH 9.0. Proteases hydrolyzed gelatin as the most specific substrate. In general, catalytic activity was efficient under a wide range of NaCl (1 to 4 M NaCl), temperature (37 to 55 °C) and after an ethanol exposition performed at -20 °C for 24 hours. In conclusion, this study reported 14 candidates extremely halophiles producing extracellular proteases capable of being stable and active on a wide range of NaCl, temperature and even long lasting ethanol exposition.

Keywords: biotechnological processes, ethanol exposition, extracellular proteases, extremophiles

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2078 Through-Bolt Moment Connection in HSS Column

Authors: Bardia Khafaf, Mehrdad Ghaffari, Amir Hussein Samakar

Abstract:

It is currently desirable to use Hollow Square Sections (HSS) in moment resistant structures in construction of building because they offer fewer restrictions for designing and more useful space while adhering to build design codes. This paper present a through bolt connection in HSS column. This connection meets building code standards that require the moment resistant connections to deflect and absorb energy resulting from gravity and seismic loads. Connection through bolts is installed and pretension to provide the connection strength needed to make a beam–column moment rigid zone. A rigid joint is typically used to resist lateral forces by holding columns and beams fixed in relation to one another. With bolted moment frames using HSS columns, a through–bolt connection could be used to secure the beam and end plate to the column. However, when multiple columns and beams are used to span a length of building, the use of through-bolts would necessities aligning multiple beams simultaneously to the columns. In the case of a linear span, the assembly process requires the holes of a first beam end plate to be aligned with through bolt holes in a column and aligning the holes of a second, opposing beam plate with the column through bolt, then inserting the through bolts in each hole for tightening with nuts and washers. In moment resistant building, a problem arises when assembling beams to columns where multiple beams and columns are required. Through bolt, moment connections are among the economical, practical and not difficult rigid steel connection for HSS column building. In this paper, the results of numerous analytical studies performed for moment structures with HSS columns with through bolt based on AISC standard codes are shown.

Keywords: through bolt, moment resistant connection, HSS columns section, construction engineering

Procedia PDF Downloads 437
2077 Bio-Detoxification of Mycotoxins by Lactic Acid Bacteria from Different Food Matrices

Authors: António Inês, Ana Guimarães, José Maria, Vânia Laranjo, Armando Venâncio, Luís Abrunhosa

Abstract:

Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP-PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 μg/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 μm pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Keywords: bio-detoxification, lactic acid bacteria, mycotoxins, food and feed

Procedia PDF Downloads 548
2076 Biodegradation of Malathion by Acinetobacter baumannii Strain AFA Isolated from Domestic Sewage in Egypt

Authors: Ahmed F. Azmy, Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

Abstract:

Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides

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2075 High-Throughput Mechanized Microfluidic Test Groundwork for Precise Microbial Genomics

Authors: Pouya Karimi, Ramin Gasemi Shayan, Parsa Sheykhzade

Abstract:

Ease shotgun DNA sequencing is changing the microbial sciences. Sequencing instruments are compelling to the point that example planning is currently the key constraining element. Here, we present a microfluidic test readiness stage that incorporates the key strides in cells to grouping library test groundwork for up to 96 examples and decreases DNA input prerequisites 100-overlay while keeping up or improving information quality. The universally useful microarchitecture we show bolsters work processes with subjective quantities of response and tidy up or catch steps. By decreasing the example amount necessities, we empowered low-input (∼10,000 cells) entire genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil miniaturized scale settlements with prevalent outcomes. We additionally utilized the upgraded throughput to succession ∼400 clinical Pseudomonas aeruginosa libraries and exhibit magnificent single-nucleotide polymorphism discovery execution that clarified phenotypically watched anti-toxin opposition. Completely coordinated lab-on-chip test arrangement beats specialized boundaries to empower more extensive organization of genomics across numerous fundamental research and translational applications.

Keywords: clinical microbiology, DNA, microbiology, microbial genomics

Procedia PDF Downloads 103